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Title: Expression of human protamine P1 in sperm of transgenic mice

Abstract

Transgenic mice were produced by pronuclear injection with DNA constructs containing human protamine P1 cDNA recombined with a murine protamine P1 promoter, and were identified by PCR. Expression of human P1 was investigated using huplm, a monoclonal antibody specific for human P1, applied to murine testicular cells, smears of epididymal sperm, and smears of detergent-isolated sperm nuclei. Various antibodies and nontransgenic littermates were used as controls. Two male founders (T3 and T7) sired more than five generations of transgenic offspring each with continued expression of human P1 in their sperm. Transgenic animals appear of normal fertility with sperm of typical nuclear morphology. The human P1 transgene was expressed postmeioticly in both lines, as expected. Nearly 100% of sperm of T3 and T7 hemizygotes labeled with huplm, consistent with complete diffusion of human P1 protein through the intercellular bridge of spermatogenic cells. Human P1 labeling of sperm nuclei was not visibly affected by sonication or by treatment with the detergent MATAB or the reducing agent DTT. A third founder female (T5) showed a transmission pattern consistent with insertion of the transgene into an X chromosome; her transgenic offspring expressed human P1 in only a small fraction of sperm. Human P1 transgenesmore » may serve as efficient targets for germinal mutations and transgenicmice may provide promising models for investigating the DNA complexes.« less

Authors:
; ; ;  [1];  [2]
  1. Lawrence Livermore National Laboratory, CA (United States)
  2. Univ. of California, Davis, CA (United States)
Publication Date:
OSTI Identifier:
88932
Report Number(s):
CONF-9405324-
Journal ID: EMMUEG; ISSN 0893-6692; TRN: 95:004220-0078
DOE Contract Number:  
W-7405-ENG-48
Resource Type:
Journal Article
Journal Name:
Environmental and Molecular Mutagenesis
Additional Journal Information:
Journal Volume: 23; Journal Issue: Suppl.23; Conference: 25. annual meeting of the Environmental Mutagen Society, Portland, OR (United States), 7-12 May 1994; Other Information: PBD: 1994
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; TRANSGENIC MICE; SPERMATOZOA; PROTAMINES; ECOLOGICAL CONCENTRATION; MUTATIONS; EVALUATION; MAN; GENES

Citation Formats

Wyrobek, A J, Keith, C, Stilwell, J, Lowe, X, and Anderson, G. Expression of human protamine P1 in sperm of transgenic mice. United States: N. p., 1994. Web.
Wyrobek, A J, Keith, C, Stilwell, J, Lowe, X, & Anderson, G. Expression of human protamine P1 in sperm of transgenic mice. United States.
Wyrobek, A J, Keith, C, Stilwell, J, Lowe, X, and Anderson, G. 1994. "Expression of human protamine P1 in sperm of transgenic mice". United States.
@article{osti_88932,
title = {Expression of human protamine P1 in sperm of transgenic mice},
author = {Wyrobek, A J and Keith, C and Stilwell, J and Lowe, X and Anderson, G},
abstractNote = {Transgenic mice were produced by pronuclear injection with DNA constructs containing human protamine P1 cDNA recombined with a murine protamine P1 promoter, and were identified by PCR. Expression of human P1 was investigated using huplm, a monoclonal antibody specific for human P1, applied to murine testicular cells, smears of epididymal sperm, and smears of detergent-isolated sperm nuclei. Various antibodies and nontransgenic littermates were used as controls. Two male founders (T3 and T7) sired more than five generations of transgenic offspring each with continued expression of human P1 in their sperm. Transgenic animals appear of normal fertility with sperm of typical nuclear morphology. The human P1 transgene was expressed postmeioticly in both lines, as expected. Nearly 100% of sperm of T3 and T7 hemizygotes labeled with huplm, consistent with complete diffusion of human P1 protein through the intercellular bridge of spermatogenic cells. Human P1 labeling of sperm nuclei was not visibly affected by sonication or by treatment with the detergent MATAB or the reducing agent DTT. A third founder female (T5) showed a transmission pattern consistent with insertion of the transgene into an X chromosome; her transgenic offspring expressed human P1 in only a small fraction of sperm. Human P1 transgenes may serve as efficient targets for germinal mutations and transgenicmice may provide promising models for investigating the DNA complexes.},
doi = {},
url = {https://www.osti.gov/biblio/88932}, journal = {Environmental and Molecular Mutagenesis},
number = Suppl.23,
volume = 23,
place = {United States},
year = {Sat Dec 31 00:00:00 EST 1994},
month = {Sat Dec 31 00:00:00 EST 1994}
}